Open Access
Open access

Expression of acid cleavable Asp-Pro linked multimeric AFP peptide in E. coli

Mollaev M., Zabolotskii A., Gorokhovets N., Nikolskaya E., Sokol M., Tsedilin A., Mollaeva M., Chirkina M., Kuvaev T., Pshenichnikova A., Yabbarov N.
Тип документаJournal Article
Дата публикации2021-10-14
Название журналаJournal of Genetic Engineering and Biotechnology
ИздательSpringer Nature
Квартиль по SCImagoQ2
Квартиль по Web of Science
Импакт-фактор 2021
ISSN1687157X, 20905920
Genetics
Biotechnology
Краткое описание
Background

Difficult to express peptides are usually produced by co-expression with fusion partners. In this case, a significant mass part of the recombinant product falls on the subsequently removed fusion partner. On the other hand, multimerization of peptides is known to improve its proteolytic stability in E. coli due to the inclusion of body formation, which is sequence specific. Thereby, the peptide itself may serve as a fusion partner and one may produce more than one mole of the desired product per mole of fusion protein. This paper proposes a method for multimeric production of a human alpha-fetoprotein fragment with optimized multimer design and processing. This fragment may further find its application in the cytotoxic drug delivery field or as an inhibitor of endogenous alpha-fetoprotein.

Results

Multimerization of the extended alpha-fetoprotein receptor-binding peptide improved its stability in E. coli, and pentamer was found to be the largest stable with the highest expression level. As high as 10 aspartate-proline bonds used to separate peptide repeats were easily hydrolyzed in optimized formic acid-based conditions with 100% multimer conversion. The major product was represented by unaltered functional alpha-fetoprotein fragment while most side-products were its formyl-Pro, formyl-Tyr, and formyl-Lys derivatives. Single-step semi-preparative RP-HPLC was enough to separate unaltered peptide from the hydrolysis mixture.

Conclusions

A recombinant peptide derived from human alpha-fetoprotein can be produced via multimerization with subsequent formic acid hydrolysis and RP-HPLC purification. The reported procedure is characterized by the lower reagent cost in comparison with enzymatic hydrolysis of peptide fusions and solid-phase synthesis. This method may be adopted for different peptide expression, especially with low amino and hydroxy side chain content.

Пристатейные ссылки: 30
Цитируется в публикациях: 2
Detecting aspartate isomerization and backbone cleavage after aspartate in intact proteins by NMR spectroscopy
Hinterholzer A., Stanojlovic V., Regl C., Huber C.G., Cabrele C., Schubert M.
Q1 Journal of Biomolecular NMR 2021 цитирований: 2
Co-Expression of a Thermally Stable and Methanol-Resistant Lipase and Its Chaperone from Burkholderia cepacia G63 in Escherichia coli
Zhang J., Tian M., Chen X., Lv P., Luo W., Wang Z., Xu J., Wang Z.
Q2 Applied Biochemistry and Biotechnology 2020 цитирований: 1
Effects of alpha-fetoprotein on the occurrence and progression of hepatocellular carcinoma
Zheng Y., Zhu M., Li M.
Q1 Journal of Cancer Research and Clinical Oncology 2020 цитирований: 17
Expression, purification, and structural analysis of the full-length human integral membrane protein γ-sarcoglycan.
Jamaleddine M., Harris M.S., Liyanage L., Cook G.A.
Q3 Protein Expression and Purification 2020 цитирований: 3
Functional reconstitution of TatB into the thylakoidal Tat translocase.
Zinecker S., Jakob M., Klösgen R.B.
Q2 Biochimica et Biophysica Acta - Molecular Cell Research 2020 цитирований: 2
Type of pH sensitive linker reveals different time-dependent intracellular localization, in vitro and in vivo efficiency in alpha-fetoprotein receptor targeted doxorubicin conjugate
Mollaev M., Gorokhovets N., Nikolskaya E., Faustova M., Zabolotsky A., Zhunina O., Sokol M., Zamulaeva I., Severin E., Yabbarov N.
Q1 International Journal of Pharmaceutics 2019 цитирований: 13
Open Access
Open access
Recombinant alpha-fetoprotein receptor-binding domain co-expression with polyglutamate tags facilitates in vivo folding in E. coli.
Mollaev M., Gorokhovets N., Nikolskaya E., Faustova M., Zabolotsky A., Sokol M., Tereshenko O., Zhunina O., Shvets V., Severin E., Yabbarov N.
Q3 Protein Expression and Purification 2018 цитирований: 4
Chemical Cleavage of an Asp-Cys Sequence Allows Efficient Production of Recombinant Peptides with an N-Terminal Cysteine Residue.
Pane K., Verrillo M., Avitabile A., Pizzo E., Varcamonti M., Zanfardino A., Di Maro A., Rega C., Amoresano A., Izzo V., Di Donato A., Cafaro V., Notomista E.
Q1 Bioconjugate Chemistry 2018 цитирований: 13
Binding characterization of the targeting drug AIMPILA to AFP receptors in human tumor xenografts
Tcherkassova J., Tsurkan S., Smirnova G., Borisova J., Moro R., Treshalina H.
Q2 Tumor Biology 2017 цитирований: 3
Open Access
Open access
Does alpha-fetoprotein contribute to the mortality and morbidity of human hepatocellular carcinoma? A commentary
Mizejewski G.J.
Q2 Journal of Hepatocellular Carcinoma 2016 цитирований: 14
Recombinant production of antimicrobial peptides in Escherichia coli: a review.
Li Y.
Q3 Protein Expression and Purification 2011 цитирований: 184
Production of recombinant antimicrobial peptides in bacteria.
Zorko M., Jerala R.
Q4 Methods in Molecular Biology 2009 цитирований: 31
Open Access
Open access
Recombinant alpha-fetoprotein C-terminal fragment: The new recombinant vector for targeted delivery
Posypanova G.A., Gorokhovets N.V., Makarov V.A., Savvateeva L.V., Kireeva N.N., Severin S.E., Severin E.S.
Q1 Journal of Drug Targeting 2008 цитирований: 24
Diagnosis of hepatocellular carcinoma.
Bialecki E.S., Di Bisceglie A.M.
Q1 HPB 2005 цитирований: 149
Analysis and control of proteolysis of recombinant proteins in Escherichia coli.
Rozkov A., Enfors S.
Q2 Advances in Biochemical Engineering/Biotechnology 2004 цитирований: 14
Метрики
Поделиться
Цитировать
ГОСТ |
Цитировать
1. Mollaev M. и др. Expression of acid cleavable Asp-Pro linked multimeric AFP peptide in E. coli // Journal of Genetic Engineering and Biotechnology. 2021. Т. 19. № 1.
RIS |
Цитировать

TY - JOUR

DO - 10.1186/s43141-021-00265-5

UR - http://dx.doi.org/10.1186/s43141-021-00265-5

TI - Expression of acid cleavable Asp-Pro linked multimeric AFP peptide in E. coli

T2 - Journal of Genetic Engineering and Biotechnology

AU - Mollaev, Murad

AU - Zabolotskii, Artur

AU - Gorokhovets, Neonila

AU - Nikolskaya, Elena

AU - Sokol, Maria

AU - Tsedilin, Andrey

AU - Mollaeva, Mariia

AU - Chirkina, Margarita

AU - Kuvaev, Timofey

AU - Pshenichnikova, Anna

AU - Yabbarov, Nikita

PY - 2021

DA - 2021/10/14

PB - Springer Science and Business Media LLC

IS - 1

VL - 19

SN - 2090-5920

ER -

BibTex |
Цитировать

@article{Mollaev_2021,

doi = {10.1186/s43141-021-00265-5},

url = {https://doi.org/10.1186%2Fs43141-021-00265-5},

year = 2021,

month = {oct},

publisher = {Springer Science and Business Media {LLC}},

volume = {19},

number = {1},

author = {Murad Mollaev and Artur Zabolotskii and Neonila Gorokhovets and Elena Nikolskaya and Maria Sokol and Andrey Tsedilin and Mariia Mollaeva and Margarita Chirkina and Timofey Kuvaev and Anna Pshenichnikova and Nikita Yabbarov},

title = {Expression of acid cleavable Asp-Pro linked multimeric {AFP} peptide in E. coli},

journal = {Journal of Genetic Engineering and Biotechnology}

}

MLA
Цитировать
Mollaev, Murad et al. “Expression of Acid Cleavable Asp-Pro Linked Multimeric AFP Peptide in E. Coli.” Journal of Genetic Engineering and Biotechnology 19.1 (2021): n. pag. Crossref. Web.